1. Unit Title: Excitable Cells Tidbit Title: Understanding membrane potential Facilitators: Thomas Torello, Harvard University Cynthia Wagner, UMBC Howard Sirotkin, Abhay Deshpande: Stony Brook University Thomas Abbott, Kristen Kimball: UCONN Jagan Srinivasan, Elizabeth Ryder: WPI Neurobiology & Physiology Group Northeast Summer Institute Stony Brook University 2013 1

2. For the Instructor: Components of the Activity Sheet 2

3. Materials required for this activity: -A large sheet of white paper (approximately 3’ x 4’). Note: this entire activity can be done with smaller activity boards. -Sticky notes (3 colors) and sizes (small and large). Prepare the activity boards: -Draw a representation of the phospholipid bilayer on the white paper across the long axis (“landscape mode”). The bilayer should divide the paper in half. -Using small stickies, prepare sticky note “ions” (Na+ and K+). These individual ions represent the ions that can move across the membrane to establish the electrical gradient). One color sticky will be K+ ions, the other will be Na+ ions (e.g. pink stickies for K+ ions and green stickies Na+ ions). Label the sticky notes (6-10 per activity board). -Using larger stickies, prepare “pools” of immovable K+ and Na+ ions (these represent the intracellular and extracellular excess concentrations of ions that maintain the chemical concentration). Ensure that the color is consistent between the big and small stickies (e.g. pink stickies for K+ ions and green stickies Na+ ions). -Using the third color stickies, label the small stickies ‘-’, and the large stickies will be ‘pools’ of – charges. These ‘-’ charges are kept deliberately vague, as they might represent Cl-, or other larger anions. -Using larger stickies, prepare Na+ and K+ channels. Ensure that the color is consistent between the big and small stickies (e.g. pink stickies for K+ channels and green stickies Na+ channels).

4. Context of our tidbit Introductory Biology Course /Applications of Physics to Biology Beginning of a Neurophysiology Section of a advanced Animal Physiology course Students are already familiar with: Cell membrane structure and function The chemical and structural nature of neurons 4

5. Tidbit: Understanding Membrane Potential To understand membrane potential, how it is generated and why it matters To recognize the interdisciplinary nature of neurobiology Goals for the Tidbit: 5

6. Objectives for the tidbit Students will be able to diagram how chemical and electrical gradients generate membrane potential 6

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8. 8 Different activities require different types of neurons… ….but all neurons need to have a baseline resting potential in order to be excitable.

9. From your pretest… Which of the following describes the ion concentrations in a typical neuron? A) [Na+] high inside, [K+] high inside B) [Na+] high outside, [K+] high outside C) [Na+] high inside, [K+] high outside D) [Na+] high outside, [K+] high inside 9

10. Set up your activity sheet using what you know about the cell 10

11. Na + K+ 11 IN OUT Na+ K+

12. Membrane Potential The potential difference across the membrane, or membrane potential (Vm), is simply the difference in charge between the inside of the cell and the outside of the cell. Potential difference is measured in volts; for cells the difference is thousandths of a volt, therefore "millivolt“ (mV) is the unit. By convention, we state the membrane potential as INSIDE relative to OUTSIDE the membrane. Thus, if Vm = -50 mV, the inside of the cell is more negatively charged than the outside. 12

13. What is the membrane potential (Vm) across this membrane? A.< 0 mV (more negative inside) B.0 mV (neutral) C.> 0 mV (more positive inside) 13

14. What could you do to change the Vm? 14

15. IN OUT K+ K + Leak channels Na + K+ 15 Na+ K+

16. IN OUT K+ What happens now ?? K + Leak channels K+ 16 Let’s start by considering only the K+ ions. K+

17. Which of these pictures is the best representation of what happens? 17 A B C No anion channels in membrane Tiny numbers of K+ cross membrane

18. 18 CHEMICAL force driving K+ out of the cell is equal to ELECTRICAL force driving K+ into the cell. This membrane potential is called the equilibrium potential for K+, or E K. Vm = E K when only K+ ion channels are present. Are K+ ions still moving?

19. So, what is the Vm across this membrane? A.< 0 mV (more negative inside) B.0 mV (Neutral) C.> 0 mV (more positive inside) 19

20. IN OUT Na+ What happens now ?? Na + 20 Now let’s look at Na+ ions. Na+ Na + Leak channels

21. What is the Vm across this membrane? A.< 0 mV (more negative inside) B.0 mV (neutral) C.> 0 mV (more positive inside) 21

22. Na+ IN OUT Na+ 22 Does your sheet look something like this? Na+ When only Na+ channels are open in the membrane, Vm = ????

23. 23 Now let’s look at a slightly more realistic representation of the resting neuron, incorporating both K+ and Na+ ions and channels

24. IN OUT K+ Na+ K+ 24 What happens now ?? Na+ K+

25. Which is closest to the real resting neuron? 25 A B C

26. Compared with a membrane containing only K+ channels, the Vm of the resting cell is A.More negative than E K B.Equal to E K C.More positive than E K 26

27. − − − − − − − − − − − − − − − − − − − − − − − - Na + K+ + + + + + + + + + + + + + + + + + + + + + + + + + + + CAPACITOR “C” BATTERY “V” volts RESISTANCE “R” Ohms CURRENT “I” − − − − − − − − − − − − − − − Basic physics in understanding cell function Membrane Capacitor circuit 27 Channel Conductance measured in ‘G’ Seimens G = 1/R

28. Summary Concentration gradients across membranes containing selective ion channels establish membrane potentials The equilibrium potential for an ion E ion occurs when the concentration gradient ‘pushing’ the ion in one direction is balanced by the electrical gradient ‘pulling’ the ion in the opposite direction The resting potential in a cell results from opposing Na+ and K+ concentration gradients, combined with the presence of both K+ and Na+ leak ion channels Cell membranes can be represented by electrical circuits 28

29. 29 In the next class, we will look at how changes in membrane potential create action potentials and how they are propagated…..

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31. Homework We talked about several situations in class today: a membrane with only K+ channels, a membrane with only Na+ channels, and a resting cell membrane with a mixture of channels. How does the membrane potential differ in each of these situations? Are Na+ and K+ ions flowing in the resting cell that has a mixture of channels? If so, in what directions? 31